TET2 as a novel epigenetic regulator for uterine function and fertility

项目摘要 不孕症和低生育力是妇女普遍存在的问题，子宫内膜适应性的失败， 植入胚胎被认为是一个重要的，但知之甚少的因素。1011 易位蛋白（TlR）是铁（II）/α-酮戊二酸（Fe（II）/α-KG）依赖性甲基胞嘧啶 双加氧酶（TET 1、TET 2和TET 3），其通过5-羟甲基化酶的反复氧化赋予活性DNA去甲基化。 甲基胞嘧啶（5 mC）至5-羟甲基胞嘧啶（5 hmC）、5-甲酰基胞嘧啶（5 fC）和5-羧基胞嘧啶 （5caC）。TcB还可以通过以下途径影响染色质修饰和基因表达，而不依赖于酶活性： 与包括转录因子（例如，WT 1）和表观遗传修饰物（例如，SIN3A）。 新出现的证据表明，泰特蛋白可能作为关键的表观遗传调节因子，在卵巢癌的发生发展中起作用。 类固醇激素雌激素（E2）和孕激素（P4）通过其同源核受体ESR 1和PGR， 正常子宫功能的调节。我们在人类和小鼠身上的初步数据表明， 在子宫内膜上皮细胞和间质细胞中表达（Tet 2>> Tet 3> Tet 1），在子宫内膜增生期间显著增加。 子宫内膜异位症在位内膜的容受窗缩短。鉴于TET 2在 女性癌症中的顶部突变基因，特别是子宫体子宫内膜癌（UCEC）和子宫内膜癌， 为了获得子宫癌肉瘤（UCS），我们产生了子宫Tet 2缺失的雌性小鼠，PgrCre/+ Tet 2f/f（Tet 2d/d）。Tet2d/d 母亲生育力低下，卵巢和输卵管功能正常。因此，本提案的目标是 使用条件性Tet 2突变小鼠模型确定Tet 2在子宫生理学中的体内作用。在目标1中， 我们将确定Tet 2在感受性窗口调节中的作用。在目标2中，我们将确定 Tet 2在体内子宫内膜间质细胞蜕膜化中的作用这些目标的完成将确定 TET 2在调节子宫内膜生理学（包括着床和蜕膜化）中的表型作用 in vivo.此外，从小鼠工作中产生的数据将为未来的标准NIH提供强有力的支持。 R 01/R21应用，解决了子宫上皮和/或间质TET 2 在怀孕和疾病状态期间调节子宫功能（例如，子宫内膜异位症和子宫内膜癌）。 因此，这项研究的结果将对人类临床医学具有重要意义。

PROJECT SUMMARY Infertility and subfertility are pervasive problems in women, and failure of the endometrial adaptation to an implanting embryo is considered a significant, yet poorly understood, contributing factor. Ten-Eleven Translocation proteins (TETs) are the iron(II)/a-ketoglutarate (Fe(II)/a-KG)-dependent methylcytosine dioxygenases (TET1, TET2 and TET3) that confer active DNA demethylation by the iterative oxidation of 5- methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC). TETs can also affect chromatin modifications and gene expression independent of enzymatic activity via interactions with other proteins including transcription factors (e.g., WT1) and epigenetic modifiers (e.g., SIN3A). Emerging evidence suggests that TET proteins may serve as key epigenetic regulators in response to ovarian steroid hormones estrogen (E2) and progesterone (P4) via their cognate nuclear receptors ESR1 and PGR in the regulation of normal uterine function. Our preliminary data in humans and mice has shown that all three TETs are expressed (Tet2>>Tet3>Tet1) in both endometrial epithelial and stromal cells, increased significantly during the window of receptivity but decreased in eutopic endometrium of endometriosis. Given that TET2 ranks among top-mutated genes in female cancers, particularly uterine corpus endometrial carcinoma (UCEC) and uterine carcinosarcoma (UCS), we have generated uterine Tet2-deleted female mice, PgrCre/+Tet2f/f (Tet2d/d). The Tet2d/d mothers were subfertile with normal ovarian and oviductal functions. Therefore, the goal of this proposal is to determine the in vivo role of Tet2 in uterine physiology using conditional Tet2 mutant mouse models. In Aim 1, we will determine the role of Tet2 in regulation of the window of receptivity. In Aim 2, we will determine the role of Tet2 in endometrial stromal cell decidualization in vivo. Completion of these aims will have defined the phenotypic roles of TET2 in the regulation of endometrial physiology, including implantation and decidualization in vivo. In addition, data generated from this mouse work will provide strong support for a future Standard NIH R01/R21 application, addressing the epigenetic mechanisms by which uterine epithelial and/or stromal TET2 regulate uterine function during pregnancy and disease states (e.g., endometriosis and endometrial cancer). Thus, the findings from this research will be important in informing human clinical medicine.